The objective of the current study was to investigate the expression pattern and clinicopathological significance of SCC-S2 in patients with non-small-cell lung cancer (NSCLC). The expression profile of SCC-S2 in NSCLC tissues and adjacent noncancerous lung tissues was detected by real-time RT-PCR, western blot analysis, and immunohistochemistry. In 25 lung cancer tissues examined, 18 (72%) of them exhibited stronger levels of SCC-S2 mRNA compared with their corresponding normal tissues. SCC-S2 protein level was up-regulated in cancerous lung tissues compared to adjacent normal tissue. Moreover, the expression level of SCC-S2 in 93 archived NSCLC tissues was measured by immunohistochemical staining. SCC-S2 was found to be overexpressed in 71 of 93 (76.3%) human lung cancer samples and correlated with lymph node metastasis (P = 0.0181), p-TNM stage (P = 0.0042), Ki-67 expression (P = 0.0028), and poor survival (P = 0.012). In addition, depleting SCC-S2 expression by small-interfering RNA inhibited growth and invasion in lung cell lines. These results indicate that SCC-S2 plays an important role in NSCLC and might be a useful therapeutic target of NSCLC.